PO.ET06.02 · 实验与分子治疗
EIK1005的临床前开发，一种强效且选择性的Werner解旋酶抑制剂
Preclinical development of EIK1005, a potent and selective inhibitor of Werner helicase
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
RecQ家族的Werner（WRN）解旋酶在DNA修复和基因组稳定性中发挥关键作用，并已被确定为微卫星不稳定性高（MSI-H）癌症中一个有前景的合成致死靶点。在此，我们呈现临床前细胞和体内数据，证明WRN抑制剂EIK1005在多种MSI-H癌症模型中具有强效且选择性的活性。EIK1005强效抑制WRN的生化活性（IC₅₀ = 30 nM），对BLM及其他相关RecQ解旋酶无活性。在MSI-H的HCT116细胞中，EIK1005诱导DNA损伤（EC₅₀ = 90 nM），并与显著的抗增殖效应相关（EC₅₀ = 28 nM）。利用我们的细胞单分子追踪平台进行的机制研究表明，EIK1005强效降低了WRN的运动，将WRN捕获在染色质上并促进WRN降解，说明了强健的靶点结合。在微卫星稳定的细胞系中未能测出EIK1005的任何效应。EIK1005具有强健的药学特性，包括在临床前物种中的高生物利用度以及剂量成比例的PK。因此，EIK1005在多种癌症小鼠模型中进行了测试。在HCT116异种移植小鼠中，EIK1005以15和30 mg/kg每日一次口服给药分别诱导了48.7%和47.6%的肿瘤消退。在对WRN抑制敏感性较低的RKO和IGROV-1 MSI-H模型中，EIK1005在所评估的30、90和180 mg/kg剂量水平上表现出剂量依赖性的肿瘤生长抑制。在HCT116和RKO肿瘤模型中观察到EIK1005强健的剂量依赖性药效学活性，分别在15和90 mg/kg的EIK1005剂量水平上观察到显著的WRN蛋白降解（给药后8小时约70%）。EIK1005在这些模型中还介导了DNA损伤应答标志物的一致升高。给予EIK1005后，在体外和小鼠血浆中均观察到GDF-15显著增加，提示GDF-15升高可能作为WRN抑制的一种外周生物标志物。在15和90 mg/kg剂量水平观察到的药效学变化与强健且剂量应答性的血浆浓度相关。进一步的研究阐明了对WRN抑制潜在耐药的若干机制。总体而言，我们对EIK1005的临床前表征证明了其强效活性和有利的药理学特征，为EIK1005在MSI-H癌症患者中正在进行的临床开发提供了动力。
查看英文原文 English abstract
The RecQ Werner (WRN) helicase plays a critical role in DNA repair and genomic stability and has been identified as a promising synthetic lethal target in microsatellite instability-high (MSI-H) cancers. Here we present preclinical cellular and in vivo data demonstrating the potent and selective activity of WRN inhibitor EIK1005 across multiple MSI-H cancer models. EIK1005 potently inhibited the biochemical activity of WRN (IC₅₀ = 30 nM), with no activity against BLM and other related RecQ helicases. In MSI-H HCT116 cells, EIK1005 induced DNA damage (EC₅₀ = 90 nM) that correlated with significant anti-proliferative effect (EC₅₀ = 28 nM). Mechanistic studies using our cellular Single Molecule Tracking platform revealed that EIK1005 potently decreased WRN motion, traps WRN on chromatin and promotes WRN degradation, illustrating robust target engagement. No effects of EIK1005 could be measured in microsatellite stable cell lines. EIK1005 has robust pharmaceutical properties including high bioavailability across preclinical species and dose proportional PK. As such, EIK1005 was tested in several mouse models of cancer. In HCT116 xenografted mice, once-daily oral dosing of EIK1005 at 15 and 30 mg/kg induced tumor regression of 48.7% and 47.6%, respectively. In RKO and IGROV-1 MSI-H models that are less sensitive to WRN inhibition, EIK1005 showed dose-dependent tumor growth inhibition across the 30, 90 and 180 mg/kg dose levels assessed. Robust dose-dependent pharmacodynamic activity of EIK1005 was observed in the HCT116 and RKO tumor models, with significant degradation of WRN protein (~ 70%, 8 h post dosing) observed at 15 and 90 mg/kg EIK1005 dose levels respectively. EIK1005 also mediated consistent elevation of DNA damage response markers in these models. A significant increase in GDF-15 was observed in vitro and in mouse plasma following EIK1005, suggesting GDF-15 elevations as a possible peripheral biomarker of WRN inhibition. The pharmacodynamic changes observed at the 15 and 90 mg/kg dose levels correlated with robust and dose responsive plasma concentrations. Further studies elucidated several mechanisms of potential resistance to WRN inhibition. Overall, our preclinical characterization of EIK1005 demonstrates its strong activity and favorable pharmacologic profile that provide impetus to the ongoing clinical development of EIK1005 in patients with MSI-H cancers.
利益披露 Disclosure
F. Rodriguez Perez, None..
D. Murnock, None..
J. Ezemba, None..
T. Ahmed, None..
J. Ortega, None..
H. M. Morrison, None..
R. Miranda, None..
T. Heuer, None..
S. Jones, None..
B. Smith, None..
A. Tabatabaei, None..
S. Basham, None..
M. Dumble, None..
M. P. Kelly, None.